JP6363104B2 - Compositions comprising 15-OHEPA and methods of using the same - Google Patents

Description

  The present invention relates to non-alcoholic fatty liver by administering a pharmaceutical composition comprising 15-hydroxyeicosapentaenoic acid (also known as 15-OHEPA or 15-HEPE) to a subject in need thereof. The present invention relates to compositions, formulations and methods for treating fatty liver disorders (FLD) such as disease (NAFLD) and non-alcoholic steatohepatitis (NASH) and their sequelae. In particular, the present invention is directed to treating subjects suffering from FLD and / or complications of FLD to reduce fat deposits in the liver to treat or prevent FLD and related complications. It relates to a pharmaceutical composition with improved efficacy against those containing EPA as a significantly active ingredient rather than -OHEPA.

  Fatty liver disorder, also known as fatty liver or fatty liver disease (FLD), is a large vesicle of triglyceride fat that enters the hepatocytes through the process of steatosis or abnormal accumulation of intracellular lipids. It relates to the accumulated state. Despite having multiple causes, fatty liver is regarded as one disease that frequently occurs in subjects with excessive alcohol intake and / or obese subjects (with or without insulin resistance effects) Yes. This condition is also associated with other diseases that affect fat metabolism. FLD can be classified into two distinct states: alcoholic FLD and non-alcoholic FLD. Both conditions show small and large lipid changes at different stages of the disease. Fat accumulation can also be accompanied by progressive inflammation of the liver (hepatitis) called steatohepatitis. Fatty liver is also known in the art as alcoholic steatosis and nonalcoholic fatty liver disease (NAFLD), with more severe forms being alcoholic steatohepatitis (part of alcoholic liver disease) and It is known as nonalcoholic steatohepatitis (NASH). Nonalcoholic fatty liver disease-related cirrhosis is the most severe form of the disease and is characterized by liver inflammation that ultimately leads to scarring of the liver tissue leading to liver failure.

  Obesity, metabolic syndrome, type 2 diabetes and atherosclerosis are increasing at a very high rate in Western countries. In recent years, fatty liver has emerged as an independent risk factor for these diseases. Fatty liver is the accumulation of triglycerides and other fats in hepatocytes. Fatty liver disease can range from fatty liver alone (also known as “liposis”) to fatty liver with inflammation or steatohepatitis. Nonalcoholic fatty liver disease (NAFLD) and nonalcoholic steatohepatitis (NASH) are the most common causes of chronic liver disease in the adult population, with significant progression to liver failure, cirrhosis and hepatocellular carcinoma Deterministic. Steatosis occurs in about 30% of the population, while 80% of obese patients have NAFLD and 50% of patients who undergo bariatric surgery have steatosis. NAFLD is also the most common cause of liver disease in children. It is estimated that NAFLD affects up to 20% of adults and nearly 5% of children. Some experts estimate that about two-thirds of obese adults and one-half of obese children can have fatty liver. Over the past decade, obesity rates have doubled in adults and tripled in children and teenagers, making NAFLD and NASH more common. NASH can cause cirrhosis, very serious disease that can require liver transplantation, and eventually scarring and sclerosis leading to hepatocellular carcinoma.

  There is no single established medical care for fatty liver. Currently, treatment of NAFLD is 1) treatment of related metabolic disorders such as diabetes and hyperlipidemia; 2) management of insulin resistance with a focus on weight loss, exercise and / or pharmacological approaches; and 3) Limited to the use of antioxidants as hepatoprotectants. Despite the use of many different therapies, no obvious treatment is currently available to address NAFLD. Since it is clinically important to resolve NAFLD and its sequelae, new approaches are needed that aim to prevent and reverse fat accumulation in the liver.

  The inventors have surprisingly found that 15-OHEPA, an EPA metabolite, is more potent than EPA in treating FLD.

  The present invention relates to fatty liver comprising non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH) by administering a composition comprising 15-OHEPA to a subject in need thereof. It relates to compositions and methods for treating disorders (FLD).

As used herein, “15-OHEPA” is 15-hydroxy-eicosa-5,8,11,13,17-pentaenoic acid. 15-OHEPA can be synthesized from eicosapentaenoic acid, EPA, by methods known in the art. As used herein, the term “15-OHEPA” refers to its free acid form of 15-OHEPA (eg, 15-hydroxy-eicosa-5,8,11,13,17-pentaenoic acid) and / or Or a pharmaceutically acceptable ester, conjugate or salt thereof, or a mixture of any of the foregoing. Derivatives of 15-OHEPA can be used instead, but this does not include any derivative compounds that lack the hydroxy group of 15-OHEPA. In some embodiments, 15-OHEPA is used in the free acid form. Alternatively, pharmaceutically acceptable esters or salts of 15-OHEPA are used in the present invention. In some embodiments, 15-OHEPA is in the form of a C _1-4 alkyl ester, such as the methyl ester or ethyl ester type.

  As used herein, “EPA” is eicosa-5,8,11,14,17-pentaenoic acid, also known as 20: 5n-3, ω-3 fatty acid. EPA can be easily obtained through commercial sources.

  Accordingly, in one aspect of the invention, there is provided a method of treating a fatty liver disorder in a subject comprising administering to the subject a therapeutically effective amount of a composition comprising 15-OHEPA.

  The present invention provides a composition comprising 15-OHEPA or 15-OHEPA for use in the treatment of fatty liver disorders.

  The present invention provides the use of 15-OHEPA or a composition comprising 15-OHEPA in the manufacture of a medicament for treating fatty liver disorders.

  In another aspect, the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of 15-OHEPA. 15-OHEPA may be the only important active ingredient in this composition as well as in the methods and uses described herein. 15-OHEPA may be the only active ingredient. Alternatively, 15-OHEPA may be combined for co-formulation or co-administration with other agents for treating FLD. If further active agents are to be used, 15-OHEPA can be co-formulated as a single dosage unit, or formulated as two or more dosage units for coordinated, combined or concomitant administration be able to.

  The invention also provides a formulation of 15-OHEPA and a formulation comprising 15-OHEPA for treating fatty liver disorders including nonalcoholic fatty liver disease (NAFLD) and nonalcoholic steatohepatitis (NASH) and Also provided are methods of using these formulations.

  15-OHEPA is a chiral molecule and can be used in the (S)-or (R) -enantiomer form or as a racemic mixture. As used herein, “15-OHEPA” includes all such types without limitation with respect to stereospecificity. In another embodiment, 15-OHEPA comprises (S) form: 15 (S) -hydroxy- (5Z, 8Z, 11Z, 13E, 17Z) -eicosapentaenoic acid. In some embodiments, 15-OHEPA can be used in the form of the ethyl ester. In other embodiments, 15-OHEPA can be used as the free acid.

  The present invention further provides a pharmaceutical composition for oral delivery comprising 15-OHEPA. The composition may include a pharmaceutically acceptable excipient. The 15-OHEPA can be any type discussed herein. 15-OHEPA may be present from about 50 mg to about 3000 mg.

  Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention is related. Although methods and materials similar or equivalent to those described herein can be used in the practice of the present invention, suitable methods and materials are described below. All publications, patent applications, patents and other references mentioned herein are expressly incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control. In addition, the materials, methods, and examples described herein are illustrative only and not intended to be limiting.

  Other features and advantages of the present invention will become apparent from the following detailed description.

and FIG. 7 presents data from in vivo efficacy studies of 15-OHEPA and EPA in a STAM model of nonalcoholic steatohepatitis, discussed in the Examples herein.

Pharmaceutical Composition While the invention may be embodied in various forms, this disclosure is to be regarded as illustrative of the invention and is not intended to limit the invention to the particular embodiments illustrated. With the understanding that it is not intended, the following description of some embodiments is provided. The headings are provided for convenience only and should not be construed as limiting the invention in any way. Embodiments shown under any heading can be combined with embodiments shown under other headings.

  Unless expressly indicated otherwise, the use of numerical values in the various quantitative values specified in this application is approximate so that the word “about” precedes both the minimum and maximum values within the stated range. As mentioned. In this way, substantially the same results as the values mentioned can be achieved using slight variations from the values mentioned. Also, the disclosure of ranges is intended as a continuous range including all values between the listed minimum and maximum values as well as any range that can be formed by such values. Also disclosed herein are any and all ratios (and any such range of ratios) that can be formed by dividing any other recited numerical value by the recited numerical value. Accordingly, those skilled in the art will be able to unambiguously obtain many such ratios, ranges and ratio ranges from the numerical values presented herein, and in all examples, such ratios, ranges and ratios. It will be appreciated that the scope of this represents various embodiments of the invention.

15-Hydroxyeicosapentaenoic acid In one embodiment, the composition of the present invention comprises 15-OHEPA as an active ingredient. 15-OHEPA is a metabolite of eicosapentaenoic acid that can be synthesized via methods known in the art, such as exposure of 15-hydroxyeicosapentaenoic acid, eicosapentaenoic acid (EPA) to the enzyme 15-lipoxygenase. Abbreviation. As used herein, the term “15-OHEPA” refers to its free acid form of 15-OHEPA (eg, 15-hydroxyeicosapentaenoic acid) and / or pharmaceutically acceptable esters, conjugates thereof. Or a salt, or a mixture of any of the foregoing. Derivatives of 15-OHEPA can be used instead, but this does not include any derivative compounds that lack the hydroxy group of 15-OHEPA. The term “pharmaceutically acceptable” in this context means that the substance does not cause unacceptable toxicity to the subject or interaction with other components of the composition.

In one embodiment, 15-OHEPA is in the form of an ester (also referred to herein as E-15-OHEPA or ethyl-15-OHEPA). In another embodiment, 15-OHEPA includes _C 1 -C ₅ alkyl esters of 15-OHEPA. In another embodiment, 15-OHEPA comprises 15-OHEPA methyl ester, 15-OHEPA propyl ester, or 15-OHEPA butyl ester. In yet another embodiment, 15-OHEPA comprises optically active 15 (S) -hydroxy- (5Z, 8Z, 11Z, 13E, 17Z) -eicosapentaenoic acid. This isomer can be used in any of the forms discussed above.

  In another embodiment, 15-OHEPA comprises lithium 15-OHEPA, mono-, di- or triglycerides of 15-OHEPA, or any other ester or salt of 15-OHEPA, or the free acid form of 15-OHEPA.

  In various embodiments, the present invention provides a pharmaceutical composition, such as an orally deliverable composition, comprising 15-OHEPA. In one embodiment, the composition comprises a therapeutically effective amount of 15-OHEPA. In one embodiment, the pharmaceutical composition comprises from about 0.1% to about 99%, from about 1% to about 95%, from about 5% to about 90% by weight of 15-OHEPA.

  In one embodiment, the pharmaceutical composition comprises about at least about 70%, at least about 80% or at least about 90% by weight of 15-OHEPA. In one embodiment, the pharmaceutical composition comprises at least about 50%, at least about 60%, at least about 70%, at least about 80% or at least about 90% by weight of 15-OHEPA.

  In another embodiment, 15-OHEPA is about 1 mg to about 10,000 mg, 25 mg to about 7500 mg, about 25 mg to about 5000 mg, about 50 mg to about 5000 mg, about 50 mg to about 3000 mg, about 75 mg to about 75 mg to about 75 mg in the composition of the invention. 2500 mg or about 100 mg to about 1000 mg, e.g., about 25 mg, about 50 mg, about 75 mg, about 100 mg, about 125 mg, about 150 mg, about 175 mg, about 200 mg, about 225 mg, about 250 mg, about 275 mg, about 300 mg, about 325 mg, about 350 mg, about 375 mg, about 400 mg, about 425 mg, about 450 mg, about 475 mg, about 500 mg, about 525 mg, about 550 mg, about 575 mg, about 600 mg, about 625 mg, about 650 mg, about 675 mg, about 700 mg, about 725 mg, about 7 0 mg, about 775 mg, about 800 mg, about 825 mg, about 850 mg, about 875 mg, about 900 mg, about 925 mg, about 950 mg, about 975 mg, about 1000 mg, about 1025 mg, about 1050 mg, about 1075 mg, about 1100 mg, about 1025 mg, about 1050 mg, About 1075 mg, about 1200 mg, about 1225 mg, about 1250 mg, about 1275 mg, about 1300 mg, about 1325 mg, about 1350 mg, about 1375 mg, about 1400 mg, about 1425 mg, about 1450 mg, about 1475 mg, about 1500 mg, about 1525 mg, about 1550 mg, about 1575 mg About 1600 mg, about 1625 mg, about 1650 mg, about 1675 mg, about 1700 mg, about 1725 mg, about 1750 mg, about 1775 mg, about 1800 mg, about 825 mg, about 1850 mg, about 1875 mg, about 1900 mg, about 1925 mg, about 1950 mg, about 1975 mg, about 2000 mg, about 2025 mg, about 2050 mg, about 2075 mg, about 2100 mg, about 2125 mg, about 2150 mg, about 2175 mg, about 2200 mg, about 2225 mg, It is present in an amount of about 2250 mg, about 2275 mg, about 2300 mg, about 2325 mg, about 2350 mg, about 2375 mg, about 2400 mg, about 2425 mg, about 2450 mg, about 2475 mg or about 2500 mg.

  In one embodiment, the 15-OHEPA present in the composition of the present invention comprises at least 90% by weight of 15-OHEPA (the term “15-OHEPA” is defined and exemplified herein). The 15-OHEPA composition can comprise higher purity 15-OHEPA, for example, at least 95% by weight 15-OHEPA or at least 97% by weight 15-OHEPA, where 15-OHEPA is indicated herein. Any form of 15-OHEPA. The purity of 15-OHEPA can be further defined by any of the 15-OHEPA descriptions provided herein (eg, a purity profile).

  Above, the amount of 15-OHEPA in the pharmaceutical composition and its purity are discussed. The nature of the essential fatty acids and their synthesis are such that the 15-OHEPA composition can include other essential fatty acid moieties in the essential fatty acid metabolism cascade.

  In one embodiment, the composition of the present invention comprises about 10% or less, about 9% or less, about 8% or less, about 7% or less, about 6% or less, about 5% or less, about 4% or less. Others containing α-linolenic acid, stearidonic acid, docosahexaenoic acid (DHA) or their derivatives in weight percent or less, about 3 weight percent or less, about 2 weight percent or less, about 1 weight percent or less, or about 0.5 weight percent or less Of omega-3 fatty acids. In other embodiments, such other omega-3 fatty acids are substantially or absent.

  In another embodiment, 15-OHEPA is at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95% of the total fatty acids present in the composition of the invention. % By weight, at least about 97% by weight, at least about 98% by weight, at least about 99% by weight or 100% by weight.

  There may be some residual eicosapentaenoic acid from the synthesis of 15-OHEPA. About 10% or less, about 9% or less, about 8% or less, about 7% or less, about 6% or less, about 5% or less, about 4% or less, about 3% or less, about 2% There may be up to about 1%, up to about 1%, or up to about 0.5% by weight of EPA. Alternatively, there is substantially or no EPA in a form that is not modified to the hydroxyl form.

Additional Active Agents In one embodiment, the pharmaceutical composition further comprises one or more additional active agents. In one embodiment, the pharmaceutical composition comprises an amount of the agent that is less than the therapeutically effective amount generally recognized for the additional active agent. In one embodiment, the pharmaceutical composition comprises an amount of the agent equal to or greater than the therapeutically effective amount generally recognized for the additional active agent.

  EPA itself has advantageous properties in the treatment of FLD, and in an alternative embodiment, 15-OHEPA can be combined with EPA.

  In one embodiment, the 15-OHEPA and one or more active agents are from about 1: 1000 to about 1000: 1, from about 1: 500 to about 500: 1, from about 1: 100 to about 100: 1, from about 1. : 50 to about 50: 1, about 1:25 to about 25: 1, about 1:10 to about 10: 1, about 1: 5 to about 5: 1, about 1: 4 to about 4: 1, about 1 Present in or co-administered in a weight ratio of 15-OHEPA: additional agent from: 3 to about 3: 1, about 1: 2 to about 2: 1 or about 1: 1.

Dosage Forms Compositions for use in accordance with the present disclosure can be formulated as one or more dosage units. The terms “dosage unit” and “dosage unit” herein refer to a portion of a pharmaceutical composition that contains a therapeutic agent in an amount suitable for a single dose to produce a therapeutic effect. Such dosage units may elicit one to multiple times per day (ie 1 to about 10, 1 to 8, 1 to 6, 1 to 4 or 1 to 2 times) or induce a therapeutic response It can be administered as many times as necessary.

  In some embodiments, the composition of the present invention is in an orally deliverable dosage form or unit form. Non-limiting examples of suitable dosage forms include tablets (eg, suspension tablets, chewable suspension tablets, rapid dispersion tablets, chewable tablets, etc.), caplets, capsules (eg, soft or hard gelatin capsules or HPMC Capsules), lozenges, sachets, cachets, troches, pellets, suspensions, elixirs, syrups, or any other solid dosage form reasonably compatible with oral administration. As used herein, the terms “oral delivery” and “oral administration” include any form of delivery in which a drug or composition is treated (in swallowed or not) into a subject's mouth. As such, this includes buccal and sublingual administration, as well as esophageal administration.

  Alternatively, the compositions of the invention can be formulated for rectal, topical or parenteral (eg, subcutaneous, intramuscular, intravenous and intradermal, or infusion) delivery.

  When discussing the amount of 15-OHEPA in the composition of the present invention, it may be divided into several dosage forms. There are restrictions on the size for oral administration. If a subject must be administered 1-4 g of 15-OHEPA per day, this can be due to up to 4 capsules each providing 1 g of 15-OHEPA.

  The composition of the present invention may be in the form of a liquid dosage form or dosage unit that is directly ingested, or may be mixed with a food or beverage prior to ingestion. Non-limiting examples of suitable liquid dosage forms include solutions, suspensions, elixirs, syrups, liquid aerosol formulations and the like.

  In another embodiment, the composition of the present invention comprises one or more pharmaceutically acceptable excipients. As used herein, the term “pharmaceutically acceptable excipient” is used as a carrier or vehicle for delivering a therapeutic agent to a subject, or to improve handling or storage properties or a unit dose of a composition. Means any substance that is added to a pharmaceutical composition to allow or promote the formation of and does not result in unacceptable toxicity or interaction with other ingredients in the composition and is not itself a therapeutic agent To do. Merely by way of example, a pharmaceutical composition according to the present disclosure comprises one or more of an antioxidant, a surfactant, a preservative, a flavoring agent, a co-solvent, a viscosity aid, a suspending aid, and a lipophilic phase. Multiple may be included.

  In one embodiment, the pharmaceutical composition comprises ascorbic acid, palmitic acid, ascorbyl palmitate, α-tocopherol, idebenone, ubiquinone, ferulic acid, coenzyme Q10, lycopene, green tea, catechin, epigallocatechin 3-gallate (EGCG) ), Green tea polyphenol (GTP), silymarin, coffee berry, resveratrol, grape seed, pomegranate extract, genisten, pycnogenol, niacinamide and the like. In one embodiment, the pharmaceutical composition comprises from about 0.01% to about 2% by weight of an antioxidant, such as about 0.01%, about 0.02%, about 0.03%, 0.04%, about 0.05%, about 0.06%, about 0.07%, about 0.08%, about 0.09%, about 0.1%, about 0% .11%, about 0.12%, about 0.13%, about 0.14%, about 0.15%, about 0.16%, about 0.17%, about 0.1% 18%, about 0.19%, about 0.2%, about 0.21%, about 0.22%, about 0.23%, about 0.24%, about 0.25 %, About 0.26%, about 0.27%, about 0.28%, about 0.29%, about 0.3%, about 0.31%, about 0.32% %, About 0.33% by weight, 0.34 wt%, about 0.35 wt%, about 0.36 wt%, about 0.37 wt%, about 0.38 wt%, about 0.39 wt%, about 0.4 wt%, about 0 .41 wt%, about 0.42 wt%, about 0.43 wt%, about 0.44 wt%, about 0.45 wt%, about 0.46 wt%, about 0.47 wt%, about 0.0. 48%, about 0.49%, about 0.5%, about 0.51%, about 0.52%, about 0.53%, about 0.54%, about 0.55 %, About 0.56%, about 0.57%, about 0.58%, about 0.59%, about 0.6%, about 0.61%, about 0.62% %, About 0.63%, about 0.64%, about 0.65%, about 0.66%, about 0.67%, about 0.68%, about 0.69% , About 0.7% by weight, about 0.7% %, About 0.72%, about 0.73%, about 0.74%, about 0.75%, about 0.76%, about 0.77%, about 0.78% %, About 0.79% by weight, about 0.8% by weight, about 0.81% by weight, about 0.82% by weight, about 0.83% by weight, about 0.84% by weight, about 0.85% by weight About 0.86 wt%, about 0.87 wt%, about 0.88 wt%, about 0.89 wt%, about 0.9 wt%, about 0.91 wt%, about 0.92 wt%, About 0.93%, about 0.94%, about 0.95%, about 0.96%, about 0.97%, about 0.98%, about 0.99%, about 1%, about 1.1%, about 1.2%, about 1.3%, about 1.4%, about 1.5%, about 1.6%, about 1.7% % By weight, about 1.8% by weight, about 1.9% by weight or Comprises about 2% by weight of one or more antioxidants.

Treatment The compositions and formulations disclosed herein can be used for the treatment of fatty liver disease. In one embodiment, the fatty liver disease is non-alcoholic fatty liver disease. In another embodiment, the fatty liver disease is nonalcoholic steatohepatitis. In one embodiment, the method comprises administering to the subject a pharmaceutical composition disclosed herein once daily, twice daily, three times daily, or four times daily.

  As used herein, “treating” or “treatment” of a disease, disorder, or condition at least in part (1) prevents the disease, disorder, or condition, ie, the disease, disorder, or condition. Preventing clinical symptoms from developing in mammals that are exposed to or susceptible to a disease, disorder or condition but have not yet experienced or experienced symptoms of the disease, disorder or condition; (2) disease, disorder Or suppress the condition, ie stop or reduce the development of the disease, disorder or condition or clinical symptoms thereof; or (3) reduce the disease, disorder or condition, ie disease, disorder or condition or clinical symptoms thereof Including causing regression. The term “prevention” for a given disease or disorder is a subject that prevents the onset of disease development if nothing has happened, is susceptible to a disorder or disease, but has not yet been diagnosed as having a disease or disorder Is meant to prevent a disease or disorder from occurring and / or to prevent further disease / disorder development if already present.

  As used herein, “effective amount” refers to the amount of active composition necessary to confer a therapeutic effect on a subject. As used herein, a “therapeutically effective amount” refers to a sufficient amount of an agent or compound to be administered that reduces, to some extent, one or more of the symptoms of the disease, disorder or condition being treated. In some embodiments, the result is a reduction and / or alleviation of a disease sign, symptom or cause, or any other desired change in a biological system. For example, in some embodiments, an “effective amount” for therapeutic use comprises a compound disclosed herein required to provide a clinically significant reduction in disease symptoms without undue adverse side effects. The amount of the composition. In some embodiments, an appropriate “effective amount” in any individual case is determined using techniques such as a dose escalation study. The term “therapeutically effective amount” includes, for example, a prophylactically effective amount. In other embodiments, an “effective amount” of a compound disclosed herein, such as a compound of formula (A) or formula (I), may produce a desired pharmacological effect or therapeutic effect without undue adverse side effects. An amount effective to achieve improvement. In other embodiments, an “effective amount” or “therapeutically effective amount” depends on changes in the subject's metabolism, age, weight, general condition, condition being treated, severity of the condition being treated, and the judgment of the prescribing physician. It is understood that it differs between subjects. The term “pharmaceutically acceptable” in this context means that the substance does not cause unacceptable toxicity to the subject or interaction with other components of the composition.

  Without further explanation, one of ordinary skill in the art will be able to use the foregoing description and the following illustrative examples to make and utilize the disclosed methods and to practice the claimed methods. The following examples are provided to facilitate the practice of the present disclosure and should not be construed as limiting the remainder of the disclosure in any way.

  The purpose of this study was to test the effects of 15-OHEPA and EPA in a STAM model of non-alcoholic steatohepatitis.

Protocol Pathogen-free 15 day pregnant C57BL / 6 mice were transferred to Charles River Laboratories in Japan Inc. (Kanagawa, Japan). NASH can be obtained by single subcutaneous injection of postnatal streptozotocin (STZ) (Sigma, USA) and feeding a high fat diet (HFD; CLEA) Japan, Japan) after 4 weeks of age (28 ± 2 days). Established in male mice. The day before the start of treatment, the mice were randomized into 5 groups of 8 mice of 5 weeks of age (35 ± 2 days). During the treatment period, the individual's body weight was measured daily and behavioral clinical signs and survival.

group:
Group 1 (vehicle): Eight NASH mice were orally administered vehicle [olive oil] in a volume of 10 mL / kg once a day from 5 to 9 weeks of age.

  Group 2 (15-OHEPA 50 mg / kg): Eight NASH mice were orally administered with a vehicle supplemented with 15-OHEPA at a dose of 50 mg / kg once a day from 5 to 9 weeks of age.

  Group 3 (15-OHEPA 500 mg / kg): Vehicles supplemented with 15-OHEPA at a dose of 500 mg / kg once daily from 5 to 9 weeks of age were administered orally to 8 NASH mice.

  Group 4 (EPA 500 mg / kg): Eight NASH mice were orally administered with a vehicle supplemented with EPA at a dose of 500 mg / kg once a day from 5 to 9 weeks of age.

  Group 5 (positive control): Eight NASH mice were orally administered pure water supplemented with telmisartan at a dose of 10 mg / kg once a day from 5 to 9 weeks of age.

At 9 weeks of age, all mice were sacrificed and the following data was recorded;
・ Individual liver weight ・ Ratio of liver weight to body weight

  Histopathological analysis of liver sections was performed using HE staining (to estimate NAFLD activity score). For HE staining, sections were excised from liver tissue paraffin blocks pre-fixed in Buan's solution and stained with Lily-Meyer hematoxylin (Muto Pure Chemicals, Japan) and eosin solution (Wako Pure Chemical Industries). NAFLD activity score (NAS) was calculated according to Kleiner's criteria (Kleiner et al., Hepatology, 2005; 41: 1313).

  Statistical tests were performed using Bonferroni's multiple comparison test. P values less than 0.05 were considered statistically significant.

Results FIG. 1.1 shows that there was no significant change in body weight in any of the experimental groups.

  FIG. 1.2 shows that only the 15-OHEPA treated group (500 mg / kg) and the positive control group (Telmisartan) have significantly reduced liver weight compared to the vehicle control.

  Figures 2.1 and 2.2 show that NAS was significantly reduced only in the 15-OHEPA (500 mg / kg) and positive control (telmisartan) groups compared to the vehicle control.

Claims (9)

  15-Hydroxy-eicosa-5,8,11,13,17-pentaenoic acid or a pharmaceutically acceptable ester, conjugate, salt, or mixture thereof (“15 -HEPE ").   The pharmaceutical composition according to claim 1, wherein the fatty liver disorder is selected from non-alcoholic fatty liver disease and non-alcoholic steatohepatitis. The 15-HEPE is, C ₁ -C ₅ containing alkyl ester, according to claim 1 or claim 2 pharmaceutical composition according.   The pharmaceutical composition according to any one of claims 1 to 3, wherein the pharmaceutical composition further comprises a pharmaceutically acceptable excipient.   The pharmaceutical composition according to any one of claims 1 to 4, wherein the pharmaceutical composition is an oral pharmaceutical composition.   6. The pharmaceutical composition according to any one of claims 1-5, wherein the 15-HEPE is present in at least about 90% by weight of the total fatty acids present in the pharmaceutical composition.   The pharmaceutical composition according to any one of claims 1 to 6, wherein other omega-3 fatty acids are substantially absent from the pharmaceutical composition.   The pharmaceutical composition according to any one of claims 1 to 7, wherein no other omega-3 fatty acids are present in the pharmaceutical composition.   9. The pharmaceutical composition according to any one of claims 1 to 8, wherein the 15-HEPE is present in the pharmaceutical composition from about 50 mg to about 3000 mg.

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